K THUT

SC K MIN DCH PHT HIN KHNG NGUYN NS1

TRONG CHN ON BNH ST XUT HUYT DENGUE

TS. Nguyn Th Kiu Anh

Ph gim c - Trung tm Y t D phng H Ni
 Mc tiu
1. Nu c ng cc bc ca quy trnh xt
nghim.
2. Thc hnh c k thut xt nghim

3. c v din gii kt qu
 NI DUNG
I. Khi nim v SXHD

II. Mt vi c im v vi rt Dengue

III. Cc k thut trong chn on vi rt Dengue

III. K thut sc k min dch trong chn on vi rt

Dengue
Khi nim v st xut huyt Dengue
Vi rt Dengue gy bnh SXHD gm 4 typ huyt
thanh c k hiu l D1, D2, D3, D4.
Biu hin bnh: st, au c, chm xut huyt,
nng c xut huyt di da, cc c quan ni
tng dn n t vong.
Bnh xut hin c tnh cht chu k
Bnh rt d ly lan v gy thnh dch
 Mt vi c im vi rt Dengue
Hnh thi: Ht vi rt hnh cu, kch thc
nh, ng knh khong 35 -50 nm
Cu trc gm:
V envelope: bn cht l lipoprotein

Trong l ARN si c trng lng 3,8 x 106

dalton
Mt vi c im vi rt Dengue (tip)
Sc khng
D b tiu dit bi cc dung mi ho tan lipid, tia
cc tm v nhit cao
Vi rt b tiu dit trong 30 pht 600C v vi gi
40C.
Trong dung dch glycerol 50 % hoc bo qun -
700 C th vi rt c th sng vi thng n vi nm
Mt vi c im vi rt Dengue (tip)
Tnh cht khng nguyn
Protein E: l 1 glycoprotein tng ng vi khng

nguyn c hiu ca tng typs vi rt Dengue

Protein mng: l 1 polypeptid tng ng vi khng

nguyn chung ca vi rt Dengue

Protein C: l khng nguyn c trng cho ton b cc

vi rt thuc nhm Flavivirus. Trong NS1 l khng

nguyn khng cu trc nm vng li ca C ca vi rt
Cc k thut trong chn on vi rt
Dengue
Chn on trc tip

Nui cy phn lp vi rt

Pht hin vt liu di truyn ca vi rt (RT-PCR)

Cc k thut trong chn on vi rt
Dengue
Chn on gin tip
K thut sc k min dch

K thut Elisa

Phn ng ngn ngng kt hng cu

Phn ng trung ho

K thut hunh quang gin tip

Phn ng kt hp b th
 K THUT
SC K MIN DCH PHT HIN
KHNG NGUYN NS1
 Mc ch

Pht hin NS1 ca vi rt Dengue trong nhim vi rt

Dengue cp
 Nguyn l
Ca s c kt qu ca dng c XN khng nguyn
Dengue NS1 bao gm 2 vch: vch T (Vch th khng
nguyn NS1), vch C (vch chng) v ging th S.
C vch C v vch T u khng nhn thy c trc
khi nh mu.
Vch C kim tra quy trnh xt nghim v lun lun
xut hin nu quy trnh xt nghim thc hin ng.
Xt nghim Dengue NS1 c th xc nh khng nguyn
virus Dengue NS1 trong mu huyt thanh, huyt tng
hoc mu ton phn vi nhy v c hiu cao.
 u im
K thut n gin, d thc hin, cho kt qu
nhanh
C th chn on sm ngay t ngy u ca
bnh
Khng i hi trang thit b
 Nhc im
Dng tnh gi: hay gp PNCT, VKDT, bnh t
min
m tnh gi: do ly mu sm

Khng phn bit c nguyn pht, th pht

 Bnh phm v bo qun

Bnh phm: Huyt thanh/huyt tng hoc mu

ton phn ly t ngy th 1 - 9 ca bnh
Bo qun bnh phm:
Mu ton phn: bo qun c 2- 3 ngy
nhit 2- 8 .
Huyt thanh hoc huyt tng: bo qun
trong khong 1 tun nhit 2-8 .
 Vt liu v trang thit b

ng nh git dng 1 ln

B kt xt nghim

T hng dn s dng
Quy trnh thc hin xt nghim
Bc 1: dng c xt nghim, mu bnh phm nhit
phng trc khi lm XN
Bc 2: Ly dng c XN ra khi ti nhm v t ln mt
kh, phng.
Bc 3: Dng ng nh git, nh 3 git mu th tng
ng vi 50 l vo ging mu (S)
Bc 3: c kt qu trong vng 15~20 pht.

Lu : khng nn c kt qu sau 20 pht.

 Din gii kt qu
m tnh: Ch 1 ng mu xut hin vng
chng C
Dng tnh: 2 ng mu xut hin vng
chng C v vng T.
Khng c gi tr: khng thy xut hin ng
mu no kt th hoc vng chng C. Phi tin
hnh lm li xt nghim vi mt thanh th khc
 Thc hnh
Anh/Ch hy din gii kt qu trng hp sau:

Trng hp 1 Trng hp 2 Trng hp 3

 Tm tt
Bc 1: dng c xt nghim, mu bnh phm
nhit phng trc khi lm XN.
Bc 2: Ly dng c XN ra khi ti nhm v t ln
mt kh, phng.
Bc 3: Dng ng nh git, nh 3 git mu th
tng ng vi 50 l vo ging mu (S).
Bc 3: c kt qu trong vng 15~20 pht.
Xin trn trng cm n!